虎眼万年青总皂苷诱导肝癌HepG-2细胞凋亡的机制研究

Study on the Apoptotic Mechanisms of Human Liver Cancer HepG-2 Cells Induced by Total Saponins of Ornithogalum caudatum

目的:探讨虎眼万年青总皂苷诱导人肝癌HepG-2细胞凋亡的机制。方法:MTT法检测虎眼万年青总皂苷对HepG-2细胞增殖的影响;倒置显微镜观察细胞形态;流式细胞仪测定细胞凋亡率、细胞线粒体膜电位和Cyt-C蛋白表达水平;透射电镜观察细胞超微结构的改变;酶标仪检测细胞内Caspase-3活性。结果:虎眼万年青总皂苷对HepG-2细胞增殖有抑制作用,IC_(50)为(79.80±0.18)μg/m L;虎眼万年青总皂苷作用细胞后,倒置显微镜下可见细胞变圆,折光性减弱,悬浮细胞增多;透射电镜下可见细胞出现典型的凋亡特征;随着虎眼万年青总皂苷浓度的增加,细胞凋亡率、细胞内Cyt-C表达水平及Caspase-3的活性均显著升高,细胞线粒体膜电位显著降低(P<0.05或P<0.01)。结论:虎眼万年青总皂苷可显著抑制人肝癌HepG-2细胞的增殖,并可通过启动线粒体途径诱导HepG-2细胞发生凋亡。

Objective： To investigate the mechanism of human liver cancer HepG-2 cells apoptosis induced by total saponins of Ornithogalum caudatum（ OCA-TS）. Methods： The anti-proliferative effects of OCA-TS on HepG-2 cells was detected by MTT assay; the inverted microscope was used to observe cell morphology; transmission electron microscope（ TEM） was used to observe the cellular ultrastructure changes; flow cytometry method was used to detect the apoptosis rate,mitochondrial membrane potential,and protein expression level of Cyt-C; Caspase-3 activity was measured by ELISA. Results： OCA-TS can inhibit the proliferation of HepG-2 cells and the IC50 was 79. 80 ± 0. 18 μg / m L. After treated by OCA-TS,cells became round,and the refractivity of cells receded,the number of suspension cells increased. By TEM method,the cells presented typical apoptosis characteristics. With the increasing of concentration of OCA-TS,cell apoptosis rate,the protein expression level of Cyt-C and the activity of Caspase-3 were increased markedly（ P〈0.05 or P〈0.01）.Conclusion： OCA-TS can effectively inhibit the proliferation of human liver cancer HepG-2 cells by inducing apoptosis of HepG-2 cells through mitochondrial pathway.